Mouse cystatin c elisa kit

Blood samples were collected and centrifuged at 10,000 rpm for 10 minutes to obtain serum. The concentration of serum neutrophil gelatinase-associated lipocalin (NGAL) and cystatin C were measured using the Mouse NGAL ELISA Kit (R&D Systems) and Mouse Cystatin C ELISA Kit (Bio Vendor R&D) according to the manufacturer's instruction.

Urine samples were collected by individually placing mice into metabolic cages (Iffa Credo, Lyon, France) for 24 h during week 10 of the study. The albumin concentration in the urine was measured by using a mouse albumin ELISA quantification kit (Bethyl Laboratories, Montgomery, TX, USA). Blood glucose, glycated haemoglobin, and systolic blood pressure were measured, as described previously [16 (link),20 (link)]. A mouse cystatin C ELISA kit (BioVendor, Brno, Czech Republic) was used to measure plasma cystatin C levels according to the manufacturer’s instructions. Mice were anesthetized via i.p. injection of sodium pentobarbital (100 mg/kg body weight; Euthatal; Sigma-Aldrich, Castle Hill, Australia) after 10 weeks (at 16 weeks of age). Both right and left kidneys were subsequently dissected, weighed and fixed in 10% formalin and embedded in paraffin as well as fresh frozen in liquid nitrogen for storage at −80 °C. Only diabetic mice with a blood glucose of ≥15 mmol/l were included in the experiments; mice with a blood glucose of <15 mmol/l and with polycystic kidneys were excluded from the study (<5% of the total number of mice).